Due to green and environment‐friendly characteristics, ultra‐high‐performance supercritical fluid chromatography has been widely used in analytical fields in recent years, but until now few reports ...are available for monosaccharide compositional analysis of macromolecule polysaccharides. In this study, an ultra‐high‐performance supercritical fluid chromatography technology with an unusual binary modifier is used to determine the monosaccharide compositions of natural polysaccharides. Each carbohydrate herein is simultaneously labeled as 1‐pheny‐3‐methyl‐5‐pyrazolone and acetyl‐derivative via pre‐column derivatizations aiming to increase UV absorption sensitivity and decrease water solubility. Ten common monosaccharides are fully separated and detected on ultra‐high‐performance supercritical fluid chromatography combined with a photo‐diode array detector by systematic optimization of multiple relevant parameters, for example, column stationary phases, organic modifiers, additives, flow rates, and so on. Compared with carbon dioxide as a mobile phase, the addition of a binary modifier increases the resolution of analytes. Additionally, this method has the advantages of small consumption of organic solvent, safety, and being environmental‐friendly. It has been successfully applied for full monosaccharide compositional analysis of heteropolysaccharides from Schisandra chinensis fruits. To sum up, a new alternative approach is provided for monosaccharide compositional analysis of natural polysaccharides.
An unprecedent glucuronoxylogalactoglucomannan (GXG′G″M), ME-2 (Mw, 2.60 × 105 g/mol; O-acetyl % = 16.7 %), was isolated and purified from water extracts of Auricularia auricula-judae (black woody ...ear). Firstly, due to much higher O-acetyl contents, we prepared its fully deacetylated products (dME-2; Mw, 2.13 × 105 g/mol) for convenient structure survey. The repeating structure-unit of dME-2 was readily proposed based on Mw determination, monosaccharide compositions, methylation analysis, free-radical degradation and 1/2D NMR spectroscopy. The dME-2 was identified as a highly branched polysaccharide with an average of 10 branches per 10 sugar backbone units. The backbone was only repeating →3)-α-Manp-(1→ residues, substituted at the C-2, C-6 and C-2,6 positions. The side chains included β-GlcAp-(1→, β-Xylp-(1→, α-Manp-(1→, α-Galp-(1→ and β-Glcp-(1→. Secondly, the complex substituted positions of O-acetyl groups in ME-2 were determined to be at C-2, C-4, C-6 and C-4,6 in the backbone and at C-2 and C-2,3 in some side chains. Finally, the anti-inflammatory activity of ME-2 was preliminarily explored on LPS-stimulated THP-1 cells. The above date not only provided the first example for structural studies of GXG′G″M type polysaccharides, but also facilitated development and application of black woody ear polysaccharides as medicinal agents or functional dietary supplements.
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The limitations of current medications for treating rheumatoid arthritis (RA) emphasize the urgent need for the development of new drugs. This study aimed to investigate the potential anti-RA ...mechanism of amygdalin using tandem mass tag (TMT)-based quantitative proteomics technology. First, the anti-RA activity of amygdalin was evaluated in a Complete Freund’s adjuvant (CFA)-induced rat model. Then, the roles and importance of proteins in the extracted rat joint tissue were evaluated using TMT-based quantitative proteomics technology. A bioinformatics analysis was used to analyze differentially abundant proteins (DAPs). A proteomics analysis identified 297 DAPs in the amygdalin group compared with the model group, of which 53 upregulated proteins and 51 downregulated proteins showed opposite regulatory trends to the DAPs produced after modeling. According to enrichment analyses of the DAPs, the signaling pathways with a high correlation degree were determined to be the complement and coagulation cascades. Furthermore, western blotting and molecular docking were used to further validate the key node proteins, e.g., complement C1s subcomponent (C1s), component C3 (C3) and kininogen 1 (Kng1). These results suggest that amygdalin may be a promising agent for treating RA by regulating the complement and coagulation cascades.
In order to establish structural-fingerprinting of polysaccharides for improvement of quality assessment, a sample preparation method based on microwave assisted free radical degradation (MFRD) of ...plant polysaccharides was proposed to produce oligosaccharides and small Mw polysaccharides. As a case study of Schisandra chinensis and S. sphenanthera fruit polysaccharides (SCP and SSP), the MFRD condition (i.e., 100 °C, 30 s and 80 W) was confirmed to be optimal. The potential structures of the MFRD products of SCP and SSP were further discussed by combinations of HILIC-ESI−-QTOF-MSE and HILIC-ESI−-Q-OT-IT-MS/MS. As followed, multivariable statistical analysis shows a clear separation of SCP and the SSP in PCA and OPLS-DA plots based HILIC-ESI−-QTOF-MSE data. The VIP plot unveils several key Q-markers (e.g., peaks 3, 8, 9, 10, 15, 25, 26, 28, 29 and 30) with significant differences and stable emergences. Furthermore, a low-polymerization compositional fingerprinting was successfully constructed for SCP and SSP using a high-performance anion-exchange chromatography with pulsed amperometric detection. Compared to the conventional sample preparation methods, the MFRD took only a few thousandth of the time to accomplish degradations of plant polysaccharides. It significantly improves sample preparations and is generally applicable to various polysaccharide samples.
•A sample preparation method based MFRD of SCP and SSP was proposed for the first time.•The potential structures of the MFRD products of SCP and SSP were discussed.•Structural-fingerprinting of SCP and SSP was built using HILIC-ESI−-QTOF-MSE and HPAEC-PAD.
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•A case study on Panax polysaccharide extracts was conducted utilizing ATR-FTIR with FD processing combined with chemometric analysis.•A nondestructive ATR-FTIR approach with FD ...processing and chemometric analysis was proposed for the first time to discriminate Panax polysaccharide extracts.•PLS and OPLS regression models are proposed to predict Panax polysaccharide extracts content.•The proposed strategy provides a nondestructive and reliable method for quality evaluation of Panax polysaccharide extracts.
Derivative spectroscopy is used to separate the small absorption peaks superimposed on the main absorption band, which is widely adopted in modern spectral analysis to increase both the valid spectral information and the identification accuracy. In this study, a method based on attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) with first-order derivative (FD) processing combined with chemometrics is proposed for rapid qualitative and quantitative analysis of Panax ginseng polysaccharides (PGP), Panax notoginseng polysaccharides (PNP), and Panax quinquefolius polysaccharides (PQP). First, ATR-FTIR with FD processing was used to establish the discriminant model combined with principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and linear discriminant analysis (LDA). After that, two-dimensional ATR-FTIR based on single-characteristic temperature as external interference (2D-sATR-FTIR) was established using ATR-FTIR with FD processing. Then, ATR-FTIR with FD processing was combined with PLS to establish and optimize the quantitative regression model. Finally, the established discriminant model and 2D-sATR-FTIR successfully distinguished PGP, PNP and PQP, and the optimal PLS regression model had a good prediction ability for the Panax polysaccharide extracts content. This strategy provides an efficient, economical and nondestructive method for the distinction and quantification of PGP, PNP and PQP in a short detection time.
Introduction
Monosaccharide compositions analysis (MCA) is indispensable for structural characterisations and structure–activity relationships of plant polysaccharides.
Objectives
To develop a ...concise and direct MCA method, we established a quantitative analysis of the multi‐monosaccharaides by single marker (QAMS) by high‐performance anion‐exchange chromatography with pulsed‐amperometric detection (HPAEC‐PAD) method.
Methodology
A stable and reproducible HPAEC‐PAD method for simultaneous determination of aldoses, ketoses and uronic acids (i.e., l‐arabinose, d‐xylose, d‐ribose, l‐rhamnose, d‐fucose, d‐mannose, d‐glucose, d‐galactose, d‐fructose, d‐glucuronic acid and d‐galacturonic acid) was established by systematic optimisation of stationary phases, column temperatures and elution programmes. On this basis, the QAMS method was proposed through comprehensive investigations of relative correction factor (RCF) variations under different influencing factors, for example, sample concentrations, flow rates, and column temperatures.
Results
Using rhamnose as an internal reference standard, the contents of the other monosaccharide components in polysaccharides from Panax quinquefolium L. and Achyranthes bidentata Bl. samples were simultaneously determined by QAMS, and there was no significant difference between the results from the QAMS and external standard method (t test, P > 0.520). In addition, a MCA fingerprinting of 30 batches of P. quinquefolium polysaccharide was established by HPAEC‐PAD, and six common peaks were assigned and determined.
Conclusions
The established HPAEC‐PAD‐QAMS method was successfully applied to the MCA of polysaccharides from P. quinquefolium and A. bidentata after optimisation of hydrolysis conditions. HPAEC‐PAD‐QAMS was proposed and established for MCA of plant polysaccharides for the first time.
This study aimed to develop a concise and direct monosaccharide compositional analysis method. A quantitative analysis of the multi‐monosaccharaides by single marker (QAMS) by high‐performance anion‐exchange chromatography with pulsed‐amperometric detection (HPAEC‐PAD) method, was proposed for the simultaneous determination of neutral and acidic monosaccharides without derivatisation. It was successfully applied to analyse the composition of monosaccharides released from Panax quinquefolium polysaccharide (PQP) and Achyranthes bidentata polysaccharide (ABP). Additionally, a monosaccharide compositional fingerprinting of PQP was established.
A modified GC-MS analytical procedure based on trimethylsilyl-dithioacetal (TMSD) derivatization has been established for a simultaneous determination of thirteen carbohydrates. Different from ...previous approaches, the current GC-MS method was featured by a powerful practicability for simultaneous detection of aldoses, uronic acids, ketoses, and amino sugars; simplifying GC-MS chromatograms and producing a single peak for each derivatized sugar, as well as high resolution, sensitivity, and repeatability. An additional liquid-liquid extraction from derivatization mixtures was performed not only to increase the detection sensitivity of amino sugars but also to decrease the by-products of derivatization. Contrarily, three amino sugars were detected at a very low intensity or not detected at all. The effect of time on monosaccharide- mercaptalated reaction was systematically investigated. The effect of trimethylsilylation on the formation of TMSD was also optimized. The established GC-MS based on TMSD derivatization was suitable for complex carbohydrate analysis and has been successfully applied for the detection of free carbohydrates in water extracts of
roots and determination of monosaccharides in
polysaccharides.
Platycodon grandiflorum (PG) has been widely applied as a conductant drug by ancient and modern traditional Chinese medicine practitioners during long-term clinical practice. However, determining how ...to guide other medicines to the targeted lungs in traditional Chinese medicine (TCM) prescription remains unclear. An ethanol soluble fraction (Fr. B) was obtained by macroporous resin and 75 % ethanol precipitate. The components were unambiguously determined as fructooligosaccharides and small molecule weight (Mw) fructans according to HILIC-ESI−-MS/MS, MS/MS and 1/2D NMR. We discovered that the Fr. B possesses the lung-oriented guidance and targetability by activating Golgi apparatus and endoplasmic reticulum (Golgi-ER) transport system. Rab21, a highly expressed transmembrane protein in the lungs, was found to be the core-affinity target of Fr. B which physically colocalized with the Golgi-ER and directly interacted with Rab21 to accelerate the uptake of extracellular therapeutic substances. The lung-oriented guidance and targetability of Fr. B was validated by the transient knockdown and overexpression of Rab21 considering dynamic observations of colocalization interactions among Fr. B, extracellular substances, and the Golgi-ER. Together, our results delineate a potential mechanism of Fr. B toward lung-oriented guidance and targetability via a direct targeting affinity of Rab21 and resulting collective stimulation of key Golgi-ER transport effectors for the acceleration of extracellular substances into the lungs.
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American ginseng (
) has long been cultivated in China for the function food and medicine. Here, ultra-high performance liquid chromatography was coupled with electrospray ionization and triple ...quadrupole mass spectrometry (UPLC-ESI
-TQ-MS) for simultaneous detection of 22 ginsenosides in American ginseng cultivated in Mudanjiang district of Heilongjiang. The extraction conditions also were optimized by a Box Behnken design experiment. The optimized result was 31.8 mL/g as ratio of liquid to raw materials, 20.3 min of extraction time, and 235.0 W of extraction powers. The quantitative MS parameters for these 22 compounds were rapidly optimized by single factor experiments employing UPLC-ESI
-multiple reaction monitoring or multiple ion monitoring (MRM/MIM) scans. Furthermore, the established UPLC-ESI
-MRM-MS method showed good linear relationships (
² > 0.99), repeatability (RSD < 3.86%), precision (RSD < 2.74%), and recovery (94⁻104%). This method determined 22 bioactive ginsenosides in different parts of the plant (main roots, hairy roots, rhizomes, leaves, and stems) and growth years (one year to four years) of
. The highest total content of the 22 analytes was in the hairy roots (1.3 × 10⁵ µg/g) followed by rhizomes (7.1 × 10⁴ µg/g), main roots (6.5 × 10⁴ µg/g), leaves (4.2 × 10⁴ µg/g), and stems (2.4 × 10⁴ µg/g). Finally, chemometric methods, hierarchical clustering analysis (HCA) and partial least squares discrimination analysis (PLS-DA), were successfully used to classify and differentiate American ginseng attributed to different growth years. The proposed UPLC-ESI
-MRM-MS coupled with HCA and PLS-DA methods was elucidated to be a simple and reliable method for quality evaluation of American ginseng.
polysaccharides have been reported to possess important activities, so quality evaluation of polysaccharides from the genus
is urgent. In this study, enzymatic digestions were performed to establish ...multiple saccharide fingerprints by ultra-performance liquid chromatography with electrospray ionization triple quadrupole linear ion trap mass spectrometry (UPLC-ESI-TQ-MS/MS) based on a multiple-reaction monitoring in negative mode. Under optimum UPLC-ESI⁻-TQ-MS/MS conditions, excellent separation and quantification of 21 constituents were achieved within 20 min on a solid core column with a 1.6 μm particle using pre-column derivatization with a PMP reagent. This method, coupled with enzymatic digestions and principal component analysis, has been successfully applied to characterize and discriminate
polysaccharides attributed to different species and plant parts. The results suggest that the proposed analytical strategy could achieve a quality evaluation of plant polysaccharides from traditional Chinese medicines.
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